Magnetic particles as affinity matrix for purification of antithrombin

Brazil MRS Meeting 2014

Mercâs, A.A.D.^1,2 ; Maciel, J.C.^3 ; Carvalho, L.B.^2

Programa de Pós-Graduação em Biologia Aplicada À Saúde, LIKA/UFPE, Cidade Universitária, Recife, PE

50670-901, Brazil^1

Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco, Cidade Universitária, Recife, PE

50670-901, Brazil^2

Universidade Federal de Roraima, Aeroporto, Boa Vista, RR

69310-000, Brazil^3

关键词: Affinity matrix;    Blood coagulation;    Coprecipitation method;    Enhanced stability;    Heparan sulfates;    Human blood plasmas;    Magnetic composites;    Magnetic particle;   

Others  :  https://iopscience.iop.org/article/10.1088/1757-899X/97/1/012002/pdf
DOI  :  10.1088/1757-899X/97/1/012002

来源: IOP

【 摘 要 】

Immobilization of biomolecules onto insoluble supports is an important tool for the fabrication of a diverse range of functional materials. It provides advantages: enhanced stability and easy separation. In this work two different magnetic composites were synthesized (MAG-PANI-HS and mDAC-HS) to human antithrombin purification. The magnetic particles (MAG) were obtained by co-precipitation method of iron salts II and III and subsequently coated with polyaniline (MAG-PANI particles). Dacron (polyethylene terephthalate) suffered a hydrazinolysis reaction to obtain a powder (Dacron hydrazide) which was subsequently magnetized (mDAC particles) also by co-precipitation method. Heparan sulfate (HS) was immobilized to MAG-PANI and mDAC retained respectively 35μg and 38.6μg per of support. The magnetic composite containing HS immobilized (MAG-PANI-HS and mDAC-HS) was incubated with human blood plasma (1mL) and then washed with NaCl gradients. Electrophoresis of proteins present in eluates showed bands of antithrombin (58kDa). A reduction in the antithrombin activity was detected in plasma that were incubated in the composites magnetic with HS immobilized, suggesting that the antithrombin was removed of the human blood plasma and then purified. Therefore, the above results suggest that both preparations: MAG-PANI-HS and mDAC-HS are able to affinity purify antithrombin, an important component of blood coagulation.